Mimura Tatsuya, Han Kyu Yeon, Onguchi Tatsuya, Chang Jin-Hong, Kim Tae-im, Kojima Takashi, Zhou Zhongjun, Azar Dimitri T
Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, IL 60612, USA.
J Vasc Res. 2009;46(6):541-50. doi: 10.1159/000226222. Epub 2009 Jun 30.
BACKGROUND/AIMS: Decorin has been shown to have antiangiogenic properties. In this study, we evaluate the involvement of membrane type 1-matrix metalloproteinase (MT1-MMP), a proangiogenic enzyme, in decorin cleavage in the cornea.
MT1-MMP expression was confirmed immunohistochemically in keratocytes and immortalized corneal fibroblast cell lines. Corneal micropockets of bFGF were used to assess the expression of decorin and MT1-MMP. Western blotting was used to evaluate decorin degradation by MT1-MMP. Aortic ring tube formation assays were used to assay the inhibitory effect of decorin and stimulatory effect of MT1-MMP on vascular endothelial cells in vitro.
We show that MT1-MMP expression is upregulated following bFGF pellet implantation in the cornea in vivo, and that MT1-MMP cleaves decorin in a time- and concentration-dependent manner in vitro. Furthermore, the addition of MT1-MMP reduces the inhibitory effects of decorin on aortic ring tube formation in vitro. Cleavage of decorin by MT1-MMP-deficient corneal cell lysates is diminished relative to that by wild-type corneal cell lysates, and an MT1-MMP knockin restores decorin processing in vitro.
The proangiogenic role of MT1-MMP in the cornea may be mediated, in part, by facilitated cleavage of corneal decorin.
背景/目的:核心蛋白聚糖已被证明具有抗血管生成特性。在本研究中,我们评估了膜型1基质金属蛋白酶(MT1-MMP)(一种促血管生成酶)在角膜中对核心蛋白聚糖的切割作用。
通过免疫组织化学法在角膜细胞和成永生化角膜成纤维细胞系中证实MT1-MMP的表达。利用碱性成纤维细胞生长因子(bFGF)的角膜微袋法评估核心蛋白聚糖和MT1-MMP的表达。采用蛋白质免疫印迹法评估MT1-MMP对核心蛋白聚糖的降解作用。利用主动脉环血管生成实验检测核心蛋白聚糖对血管内皮细胞的抑制作用以及MT1-MMP的刺激作用。
我们发现,在体内,bFGF颗粒植入角膜后MT1-MMP的表达上调,并且在体外MT1-MMP以时间和浓度依赖性方式切割核心蛋白聚糖。此外,添加MT1-MMP可降低核心蛋白聚糖在体外对主动脉环血管生成的抑制作用。与野生型角膜细胞裂解液相比,MT1-MMP缺陷型角膜细胞裂解液对核心蛋白聚糖的切割作用减弱,并且MT1-MMP基因敲入可在体外恢复核心蛋白聚糖的加工过程。
MT1-MMP在角膜中的促血管生成作用可能部分是通过促进角膜核心蛋白聚糖的切割来介导的。